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目的筛选鉴定抗登革1~4型病毒单克隆抗体,并对其亚型、免疫学交叉反应及病毒中和能力进行初步分析。方法登革1~4型病毒乳鼠脑悬液混合后免疫BALB/c小鼠,经细胞融合后,免疫荧光法筛选抗登革病毒单克隆抗体,并在Hela细胞水平,病毒中和实验鉴定其对登革1型、2型病毒的中和能力,并详细分析所获中和抗体的亚型及免疫学交叉反应性。结果筛选获得抗登革病毒单克隆抗体41株,初步的中和实验结果表明,其中7株具登革1型病毒中和能力,13株具登革2型病毒中和能力,分别属于Ig G1、Ig G3、Ig G2a、Ig G2b、Ig M亚型,免疫荧光法交叉反应分析表明与登革1型病毒、登革2型病毒、登革3型病毒、登革4型病毒、乙型脑炎病毒、黄热病毒存在广泛免疫学交叉反应。结论筛选鉴定系列登革病毒中和抗体,为进一步探讨其中和作用的分子机制奠定了基础。
Objective To screen and identify anti-dengue monoclonal antibodies against type 1 ~ 4 viruses and to analyze their subtype, immunological cross-reaction and virus neutralization ability. Methods The BALB / c mice were immunized with the brain suspension of dengue virus type 1 ~ 4 virus. After fusion, the monoclonal antibodies against Dengue virus were screened by immunofluorescence. The results of Hela cells and virus neutralization assay Its dengue type 1, type 2 virus neutralization capacity, and detailed analysis of the subtype and immunological cross-reactivity of neutralizing antibodies. Results 41 strains of monoclonal antibodies against Dengue virus were screened. The preliminary neutralization results showed that 7 of them were capable of neutralizing Dengue virus type 1 and 13 of Dengue virus type 2 neutralizing capacity, belonging to Ig G1 , Ig G3, Ig G2a, Ig G2b, Ig M subtypes. Immunofluorescence cross-reactivity analysis showed that there was significant difference between dengue virus type 1, dengue virus type 2, dengue virus type 3, dengue virus type 4, There are a wide range of immunological cross-reactions between the inflammatory and yellow fever viruses. Conclusion Screening and identification of neutralizing antibodies against dengue virus provide a basis for further investigation on the molecular mechanism of neutralization.